Redox environment in stem and differentiated cells: A quantitative approach

Stem cells are believed to maintain a specific intracellular redox status through a combination of enhanced removal capacity and limited production of ROS. In the present study, we challenge this assumption by developing a quantitative approach for the analysis of the pro- and antioxidant ability of...

Full description

Bibliographic Details
Main Authors: O.G. Lyublinskaya, Ju.S. Ivanova, N.A. Pugovkina, I.V. Kozhukharova, Z.V. Kovaleva, A.N. Shatrova, N.D. Aksenov, V.V. Zenin, Yu.A. Kaulin, I.A. Gamaley, N.N. Nikolsky
Format: Article
Language:English
Published: Elsevier 2017-08-01
Series:Redox Biology
Online Access:http://www.sciencedirect.com/science/article/pii/S2213231717301775
id doaj-05a41ce2b7a84595905fc35d4985e38e
record_format Article
spelling doaj-05a41ce2b7a84595905fc35d4985e38e2020-11-25T01:31:28ZengElsevierRedox Biology2213-23172017-08-0112758769Redox environment in stem and differentiated cells: A quantitative approachO.G. Lyublinskaya0Ju.S. Ivanova1N.A. Pugovkina2I.V. Kozhukharova3Z.V. Kovaleva4A.N. Shatrova5N.D. Aksenov6V.V. Zenin7Yu.A. Kaulin8I.A. Gamaley9N.N. Nikolsky10Department of Intracellular Signaling and Transport, Institute of Cytology, Russian Academy of Sciences, Tikhoretsky pr. 4, St. Petersburg 194064, Russia; Corresponding author.Department of Intracellular Signaling and Transport, Institute of Cytology, Russian Academy of Sciences, Tikhoretsky pr. 4, St. Petersburg 194064, Russia; Department of Medical Physics, Institute of Physics, Nanotechnology and Telecommunications, Peter the Great St. Petersburg Polytechnic University, Polytechnicheskaya st. 29, St. Petersburg 195251, RussiaDepartment of Intracellular Signaling and Transport, Institute of Cytology, Russian Academy of Sciences, Tikhoretsky pr. 4, St. Petersburg 194064, RussiaDepartment of Intracellular Signaling and Transport, Institute of Cytology, Russian Academy of Sciences, Tikhoretsky pr. 4, St. Petersburg 194064, RussiaDepartment of Intracellular Signaling and Transport, Institute of Cytology, Russian Academy of Sciences, Tikhoretsky pr. 4, St. Petersburg 194064, RussiaDepartment of Intracellular Signaling and Transport, Institute of Cytology, Russian Academy of Sciences, Tikhoretsky pr. 4, St. Petersburg 194064, RussiaDepartment of Intracellular Signaling and Transport, Institute of Cytology, Russian Academy of Sciences, Tikhoretsky pr. 4, St. Petersburg 194064, RussiaDepartment of Intracellular Signaling and Transport, Institute of Cytology, Russian Academy of Sciences, Tikhoretsky pr. 4, St. Petersburg 194064, RussiaDepartment of Intracellular Signaling and Transport, Institute of Cytology, Russian Academy of Sciences, Tikhoretsky pr. 4, St. Petersburg 194064, RussiaDepartment of Intracellular Signaling and Transport, Institute of Cytology, Russian Academy of Sciences, Tikhoretsky pr. 4, St. Petersburg 194064, RussiaDepartment of Intracellular Signaling and Transport, Institute of Cytology, Russian Academy of Sciences, Tikhoretsky pr. 4, St. Petersburg 194064, RussiaStem cells are believed to maintain a specific intracellular redox status through a combination of enhanced removal capacity and limited production of ROS. In the present study, we challenge this assumption by developing a quantitative approach for the analysis of the pro- and antioxidant ability of human embryonic stem cells in comparison with their differentiated descendants, as well as adult stem and non-stem cells. Our measurements showed that embryonic stem cells are characterized by low ROS level, low rate of extracellular hydrogen peroxide removal and low threshold for peroxide-induced cytotoxicity. However, biochemical normalization of these parameters to cell volume/protein leads to matching of normalized values in stem and differentiated cells and shows that tested in the present study cells (human embryonic stem cells and their fibroblast-like progenies, adult mesenchymal stem cells, lymphocytes, HeLa) maintain similar intracellular redox status. Based on these observations, we propose to use ROS concentration averaged over the cell volume instead of ROS level as a measure of intracellular redox balance. We show that attempts to use ROS level for comparative analysis of redox status of morphologically different cells could lead to false conclusions. Methods for the assessment of ROS concentration based on flow cytometry analysis with the use of H2DCFDA dye and HyPer, genetically encoded probe for hydrogen peroxide, are discussed. Keywords: Embryonic stem cells, Differentiated cells, ROS, Redox status, H2DCFDA, HyPer, Flow cytometry, Quantitative redox biologyhttp://www.sciencedirect.com/science/article/pii/S2213231717301775
collection DOAJ
language English
format Article
sources DOAJ
author O.G. Lyublinskaya
Ju.S. Ivanova
N.A. Pugovkina
I.V. Kozhukharova
Z.V. Kovaleva
A.N. Shatrova
N.D. Aksenov
V.V. Zenin
Yu.A. Kaulin
I.A. Gamaley
N.N. Nikolsky
spellingShingle O.G. Lyublinskaya
Ju.S. Ivanova
N.A. Pugovkina
I.V. Kozhukharova
Z.V. Kovaleva
A.N. Shatrova
N.D. Aksenov
V.V. Zenin
Yu.A. Kaulin
I.A. Gamaley
N.N. Nikolsky
Redox environment in stem and differentiated cells: A quantitative approach
Redox Biology
author_facet O.G. Lyublinskaya
Ju.S. Ivanova
N.A. Pugovkina
I.V. Kozhukharova
Z.V. Kovaleva
A.N. Shatrova
N.D. Aksenov
V.V. Zenin
Yu.A. Kaulin
I.A. Gamaley
N.N. Nikolsky
author_sort O.G. Lyublinskaya
title Redox environment in stem and differentiated cells: A quantitative approach
title_short Redox environment in stem and differentiated cells: A quantitative approach
title_full Redox environment in stem and differentiated cells: A quantitative approach
title_fullStr Redox environment in stem and differentiated cells: A quantitative approach
title_full_unstemmed Redox environment in stem and differentiated cells: A quantitative approach
title_sort redox environment in stem and differentiated cells: a quantitative approach
publisher Elsevier
series Redox Biology
issn 2213-2317
publishDate 2017-08-01
description Stem cells are believed to maintain a specific intracellular redox status through a combination of enhanced removal capacity and limited production of ROS. In the present study, we challenge this assumption by developing a quantitative approach for the analysis of the pro- and antioxidant ability of human embryonic stem cells in comparison with their differentiated descendants, as well as adult stem and non-stem cells. Our measurements showed that embryonic stem cells are characterized by low ROS level, low rate of extracellular hydrogen peroxide removal and low threshold for peroxide-induced cytotoxicity. However, biochemical normalization of these parameters to cell volume/protein leads to matching of normalized values in stem and differentiated cells and shows that tested in the present study cells (human embryonic stem cells and their fibroblast-like progenies, adult mesenchymal stem cells, lymphocytes, HeLa) maintain similar intracellular redox status. Based on these observations, we propose to use ROS concentration averaged over the cell volume instead of ROS level as a measure of intracellular redox balance. We show that attempts to use ROS level for comparative analysis of redox status of morphologically different cells could lead to false conclusions. Methods for the assessment of ROS concentration based on flow cytometry analysis with the use of H2DCFDA dye and HyPer, genetically encoded probe for hydrogen peroxide, are discussed. Keywords: Embryonic stem cells, Differentiated cells, ROS, Redox status, H2DCFDA, HyPer, Flow cytometry, Quantitative redox biology
url http://www.sciencedirect.com/science/article/pii/S2213231717301775
work_keys_str_mv AT oglyublinskaya redoxenvironmentinstemanddifferentiatedcellsaquantitativeapproach
AT jusivanova redoxenvironmentinstemanddifferentiatedcellsaquantitativeapproach
AT napugovkina redoxenvironmentinstemanddifferentiatedcellsaquantitativeapproach
AT ivkozhukharova redoxenvironmentinstemanddifferentiatedcellsaquantitativeapproach
AT zvkovaleva redoxenvironmentinstemanddifferentiatedcellsaquantitativeapproach
AT anshatrova redoxenvironmentinstemanddifferentiatedcellsaquantitativeapproach
AT ndaksenov redoxenvironmentinstemanddifferentiatedcellsaquantitativeapproach
AT vvzenin redoxenvironmentinstemanddifferentiatedcellsaquantitativeapproach
AT yuakaulin redoxenvironmentinstemanddifferentiatedcellsaquantitativeapproach
AT iagamaley redoxenvironmentinstemanddifferentiatedcellsaquantitativeapproach
AT nnnikolsky redoxenvironmentinstemanddifferentiatedcellsaquantitativeapproach
_version_ 1725086485820997632